改良体外光化学疗法提高淋巴细胞凋亡诱导率  

The modified extracorporeal photochemotherapy promotes apoptosis of spleen lymphocytes

作　　者：周林[1,2] 杜国盛[2] 郑德华[2] 刘志佳[3] 魏玉香[3] 陈文[1] 朱志东[2] 宋继勇[2] 付晓倩[2] 

机构地区：[1]解放军总医院肝胆外科,北京100853 [2]解放军第309医院全军器官移植研究所肝胆外科,北京100091 [3]解放军第309医院全军器官移植研究所泌尿一科,北京100091

出　　处：《细胞与分子免疫学杂志》2014年第10期1099-1102,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(811022422);首都临床特色应用研究(Z111107058811008)

摘　　要：目的 探讨改良体外光化学治疗（ECP）方法提高淋巴细胞凋亡诱导率的效果。方法 以无菌离体移植肝供者脾脏作为实验材料,分别以改良和传统ECP方法制备供者脾脏淋巴细胞悬液（SP）,所得SP经8-甲氧基补骨脂素（8-MOP）联合A波段长波紫外线（UVA）照射（PUVA）、8-MOP、UVA处理,并分别设对照组;不同处理的SP于37℃、50 mL/L CO2孵箱中培养过夜（6-8 h）,通过观察细胞形态学改变并用流式细胞术检测凋亡率,比较各组间淋巴细胞凋亡率的差异。结果 改良ECP方法制备的SP,经PUVA、8-MOP、UVA处理早期凋亡率和总凋亡率分别为（95.33±3.03）%和（97.10±2.12）%,（23.39±4.55）%和（36.32±6.63）%,（66.98±3.60）%和（68.65±4.35）%,与对照组（12.82±1.86%和13.4±2.65%）相比,差异显著,具有统计学意义（P〈0.01）;传统ECP法各实验组早期及总凋亡率分别为（79.73±4.21）%和（82.70±4.13）%,（61.42±2.28）%和（68.91±2.18）%,（19.30±1.78）%和（28.06±1.88）%,（10.84±0.98）%和（12.77±1.22）%,组间比较差异显著,具有统计学意义（P〈0.01）;改良法与传统ECP法相比,早期和总凋亡率有显著性差异（P〈0.01）,而晚期凋亡率无显著性差异（P〉0.05）。结论 改良型ECP方法可简便、安全、高效提高离体脾脏淋巴细胞早期凋亡率,为进一步研究ECP诱导的树突细胞免疫调节作用奠定基础。Objective To explore the efficacy of the modified extracorporeal photochemotherapy（ ECP） in improving the apoptotic rate of lymphocytes in vitro. Methods The spleens which were obtained from liver transplantation donor under aseptic condition were used as experimental materials. Splenic lymphocytes（ SPs） suspensions were prepared by modified and traditional ECP method,respectively. And then the isolated SPs were treated by the irradiation of 8-methoxypsoralen（ 8-MOP） combined with ultraviolet A（ UVA） named PUVA,8-MOP and UVA,and compared with a blank group meanwhile.The treated SPs were cultured overnight in an incubator at 37℃,in a humidified atmosphere of 50 mL /L CO2 for 6-8 hours.The morphological changes of cells were observed using an inverted microscope,the apoptotic rates of SPs were detected by flow cytometry,and the difference between groups was analyzed finally. Results The apoptotic rate at early stage and the total apoptotic rate of SPs prepared by the modified ECP method were respectively（ 95. 33 ± 3. 03） % and（ 97. 10 ±2. 12） % after treated by PUVA,（ 23. 39 ± 4. 55） % and（ 36. 32 ± 6. 63） % after treated by 8-MOP,and（ 66. 98 ± 3. 60） % and（ 68.65 ± 4.35） % by UVA. Compared with control group（ 12. 82 ± 1. 86% and 13. 4 ± 2. 65%）,there were statistically significant differences（ P 〈0.01）. The apoptotic rate at early stage and the total apoptotic rate of SPs prepared by the traditional ECP method were respectively（ 79. 73 ± 4. 21） % and（ 82. 70 ± 4. 13） %,（ 61. 42 ± 2. 28） % and（ 68. 91 ± 2. 18） %,（ 19. 30 ±1. 78） % and（ 28. 06 ± 1. 88） %,（ 10. 84 ± 0. 98） % and（ 12. 77 ± 1. 22） %,and the statistical comparisons between groups also had significant difference（ P〈0. 01）. In addition,there was a significant difference in the early and total apoptosis between the modified and traditional ECP（ P〈0. 01）,but no obvious variation in the end-stage apoptosis in the two groups（ P 0.05）. Conclusion

关 键 词：改良 体外光化学治疗 淋巴细胞凋亡 树突状细胞 

分 类 号：R392.11[医药卫生—免疫学] R733.61[医药卫生—基础医学]